1200 series high performance liquid chromatography

Manufactured by Agilent Technologies

Sourced in United States

The 1200 series high-performance liquid chromatography (HPLC) is a analytical instrumentation designed for the separation, identification, and quantification of various chemical compounds. It utilizes a liquid mobile phase to separate the components of a mixture and detect them using various detection methods.

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Lab products found in correlation

P 2000 polarimeter, by Jasco (3 mentions) 6130 quadrupole mass spectrometer, by Agilent Technologies (2 mentions) Jms 700 high resolution mass spectrometer, by JEOL (1 mentions) Avance 600 mhz spectrometer, by Bruker (1 mentions) 321 hplc pump, by Gilson (1 mentions) 151 uv vis detector, by Gilson (1 mentions) 3000 a micro gas chromatograph, by Agilent Technologies (1 mentions) 6130 quadrupole ms, by Agilent Technologies (1 mentions) Ft ir 4200 spectrometer, by Jasco (1 mentions) Chirascan plus spectrometer, by Applied Photophysics (1 mentions) Avance 800 mhz, by Bruker (1 mentions) 400 mhz, by JEOL (1 mentions) Chirascan plus cd spectrometer, by Applied Photophysics (1 mentions) Acrylamide monomer, by Merck Group (1 mentions) Toc vcph analyzer, by Shimadzu (1 mentions) Optima 5300 dv, by PerkinElmer (1 mentions) Millipore filter, by Merck Group (1 mentions) Hplc system, by Agilent Technologies (1 mentions) Tsk gel g2000 swxl column, by Tosoh (1 mentions) Pursuit xrs ultra c18 column, by Agilent Technologies (1 mentions)

10 protocols using 1200 series high performance liquid chromatography

NMR and Mass Spectrometry Analysis

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¹H, ¹³C, and 2D (COSY, HSQC, HMBC) nuclear magnetic resonance (NMR) spectra were measured in methanol-d₄ using Bruker Avance 600 MHz spectrometers (Bruker, Billerica, MA, USA) at the National Instrumentation Center for Environmental Management (NICEM) of Seoul National University. Liquid chromatography/mass spectrometry (LC/MS) data were obtained with an Agilent Technologies 6130 quadrupole mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) coupled with an Agilent Technologies 1200 series high-performance liquid chromatography (HPLC) instrument. High-resolution fast atom bombardment (HR-FAB) mass spectra were recorded on a Jeol JMS-700 high-resolution mass spectrometer (Jeol, Tokyo, Japan) at the National Center for Inter-University Research Facilities (NCIRF) of Seoul National University. HPLC was performed using a Gilson 321 HPLC pump with a Gilson UV/VIS-151 detector (Gilson, Middleton, WI, USA). All solvents used were of spectroscopic grade or were distilled prior to use.

Chung B., Kwon O.S., Shin J, & Oh K.B. (2020). Antibacterial Activity and Mode of Action of Lactoquinomycin A from Streptomyces bacillaris. Marine Drugs, 19(1), 7.

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Gas and Metabolite Analysis in Cultures

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N₂O, CO₂, and H₂ were analyzed by manually injecting 100 µL headspace samples into an Agilent 3000 A Micro-Gas Chromatograph (Palo Alto, CA, USA) equipped with Plot Q and molecular sieve columns coupled with a thermal conductivity detector^{41 (link)}. Aqueous concentrations (µM) were calculated from the headspace partial pressures based on reported Henry’s law constants^{113 (link)} for N₂O (2.4 × 10⁻⁴), H₂ (7.8 × 10⁻⁶) and CO₂ (3.3 × 10⁻⁴) mol (m³ Pa)⁻¹ according to

H^{c p} R T = \frac{C_{a}}{C_{g}}

Where H^cp is the Henry’s law constant^{113 (link)}, R is the universal gas constant, T is the temperature, C_g is the headspace gas-phase concentration, and C_aq is the liquid phase (dissolved) concentration. Five-point standard curves for N₂O, CO₂ and H₂ spanned concentration ranges of 8333 to 133,333 ppmv. Pyruvate, acetate and formate were measured with an Agilent 1200 Series high-performance liquid chromatography (HPLC) system (Palo Alto, CA, USA)^{41 (link)}. pH was measured in 0.4 mL samples of culture supernatant following removal of cells by centrifugation with a calibrated pH electrode.

He G., Chen G., Xie Y., Swift C.M., Ramirez D., Cha G., Konstantinidis K.T., Radosevich M, & Löffler F.E. (2024). Sustained bacterial N2O reduction at acidic pH. Nature Communications, 15, 4092.

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Spectroscopic Analysis of Molecular Compounds

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Specific rotations were obtained using a JASCO P-2000 polarimeter with a 1-cm cell at 25°C. UV spectra were obtained using an Applied Photophysics Chirascan™-plus spectrometer with a 1-cm quartz cell at 25°C. IR spectral data were obtained using a JASCO FT/IR-4200 spectrometer. NMR spectra were recorded on an 800 MHz Bunker Avance III HD spectrometer with a 5-mm TCI cryoprobe and a Bunker Avance 600 MHz spectrometer at the National Center for Inter-university Research Facilities (NCIRF). LC-MS and low-resolution electrospray ionization mass spectroscopic (LR-ESI-MS) data were obtained using an Agilent Technologies 1200 series high performance liquid chromatography (HPLC) coupled with an Agilent Technologies 6130 quadrupole MS. High-resolution fast atom bombardment MS (HR-FAB-MS) data were obtained using a JEOL JMS-700 high-resolution MS at NCIRF.

Shin D., Byun W.S., Moon K., Kwon Y., Bae M., Um S., Lee S.K, & Oh D.C. (2018). Coculture of Marine Streptomyces sp. With Bacillus sp. Produces a New Piperazic Acid-Bearing Cyclic Peptide. Frontiers in Chemistry, 6, 498.

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Spectroscopic Characterization of Compounds

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Optical A JASCO P-2000 polarimeter (sodium light source, JASCO, Easton, PA, USA) with 1 cm cell was used to measure optical rotation data. Applied Photophysics Chirascan-plus CD spectrometer (Applied Photophysics, Leatherhead, Surrey, UK) with 1 mm and 2 mm CD cells was utilized to collect ultraviolet (UV) and circular dichroism (CD) spectroscopic data. A JASCO FT/IR-4200 spectrometer (FT/IR-4200, Tokyo, Japan) was used to acquire infrared (IR) spectra. NMR spectra were collected at Seoul National University’s College of Pharmacy, Seoul, Republic of Korea, using Bruker Avance 800 MHz (Bruker, Billerica, MA, USA) and JEOL 400 MHz (JEOL, Tokyo, Japan) spectrometers. An Agilent Technologies 6130 Quadrupole mass spectrometer coupled with an Agilent Technologies 1200 series high-performance liquid chromatography (HPLC) apparatus (Agilent Technologies, Santa Clara, CA, USA) enabled low-resolution electrospray ionization mass spectroscopic (LR-ESI-MS) data to be obtained. High-resolution electrospray ionization mass spectroscopic (HR-ESI-MS) data were collected in the National Instrumentation Center for Environmental Management (NICEM) at the College of Agriculture and Life Sciences at Seoul National University using an AB SCIEX Q-TOF 5600 HR-MS spectrometer (Framingham, MA, USA).

Huynh T.H., Bae E.S., Heo B.E., Lee J., An J.S., Kwon Y., Nam S.J., Oh K.B., Jang J., Lee S.K, & Oh D.C. (2023). Tandocyclinones A and B, Ether Bridged C-Glycosyl Benz[a]anthracenes from an Intertidal Zone Streptomyces sp.. Marine Drugs, 21(9), 500.

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Quantification of Acrylamide Monomer Release

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Three circular gels (1.5mm thick, 3mm diameter) per time point were placed in a 24 well plate with 2 mL 1X PBS media to extract the residual monomer from the gels. Samples were aliquot and analyzed after 30min, 1 h, 1 d, 2 d and 6 d. A standard curve of acrylamide monomer (Sigma) in 1X PBS was prepared by serial dilutions and run on 0.1% formic acid as the mobile phase. An Agilent 1200-series high-performance liquid chromatography (HPLC) coupled with a mass spectrometer was used for detection.^{[44 ]} Between runs, the machine was flushed, and the conditioned PBS was run to compare the integrated peak area in the total ion count channel of the sample to those of the known concentrations from the standard curve.

Freedman B.R., Uzun O., Luna N.M., Rock A., Clifford C., Stoler E., Östlund-Sholars G., Johnson C, & Mooney D.J. (2021). Degradable and Removable Tough Adhesive Hydrogels. Advanced materials (Deerfield Beach, Fla.), 33(17), e2008553.

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HPLC Analysis of TCH Samples

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The TCH samples were filtered through a Millipore filter (0.22 μm), and then analyzed by means of a 1200 series high-performance liquid chromatography (HPLC, Agilent, USA) with a UV detector at 365 nm and XDB-C18 column (4.6 × 150 mm, 5 μm). The mobile phase was acetonitrile and 0.01 mol L⁻¹ aqueous oxalic acid (31 : 69, v/v) mixture at room temperature, with a constant flow rate of 1.0 mL min⁻¹. The gas ozone concentration was measured by the iodometric method.^{21 (link)} The dissolved ozone concentration in water was measured by the indigo method.^{22 (link)}After each reuse, the CoSO catalyst was washed with deionized water several times and dried 24 h at 100 °C. The released cobalt ion was measured by inductively coupled plasma (ICP) on an OPTIMA 5300DV (PerkinElmer Co., USA). Total organic carbon (TOC) was determined by a Shimadzu TOC-V_CPH analyzer. The point of zero charge (pH_pzc) of CoSO catalyst was measured with a Zetasizer Nano (Malvern, UK).

Luo L., Zou D., Lu D., Yu F., Xin B, & Ma J. (2018). Study of catalytic ozonation for tetracycline hydrochloride degradation in water by silicate ore supported Co3O4. RSC Advances, 8(72), 41109-41116.

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Protein Aggregation Stability Measurement

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Protein aggregation was measured using SEC in storage stability studies with triplicate samples. The powders were weighed (2 − 4 mg per vial), sealed and stored in an oven at 40°C. The vials of each formulation were removed from the oven at different time points (15, 30, 60 and 90 days) and reconstituted to obtain a protein concentration of 1 mg/mL. To remove any insoluble aggregates prior to SEC analysis, the reconstituted solutions were centrifuged at 12,000 rpm for 10 min at 4°C and the supernatant collected for analysis. A 1200 series high performance liquid chromatography (HPLC) system (Agilent Technologies, Santa Clara, CA) with a TSK gel G2000SWXL column (Tosoh Bioscience LLC, King of Prussia, PA) operating isocratically for 16 min at a flow rate of 1mL/min was used to analyze the samples. A mobile phase of sodium phosphate buffer (pH 6.8) was used. The physical instability of the samples was determined by calculating the percentage loss of area under the curve for the monomeric peak.

Mutukuri T.T., Wilson N.E., Taylor L.S., Topp E.M, & Zhou Q.T. (2020). Effects of Drying Method and Excipient on the Structure and Physical Stability of Protein Solids: Freeze Drying vs. Spray Freeze Drying. International journal of pharmaceutics, 594, 120169.

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Characterization of Oba01, an ADC Targeting DR5

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The fully humanized DR5-specific monoclonal antibody (zaptuzumab)²⁷ (link)^,³⁰ (link) and its ADC (Oba01) were generated by Yantai Mabplex International Bio-Pharmaceutical (Shandong, China).³⁷ (link) Oba01 was produced by enabling zaptuzumab coupled with a highly toxic inhibitor of tubulin, MMAE, via a proteinase cleavable linker by ThioBridge technology and formulated in 5 mM histidine, 175 mM trehalose, and 0.035% (w/v) Tween 20 (pH 6.0). The solution was filtered (0.22 μm) and stored at −80°C.³⁷ (link)
Characterization of Oba01 was performed by size-exclusion chromatography (1200 series high-performance liquid chromatography [HPLC], Agilent Technologies, Wilmington, DE, USA) and hydrophobic interaction chromatography (HIC) as described by Hamblett et al.³⁷ (link)^,³⁸ (link) The identification for peaks corresponding to ADCs with 2, 4, and 6 mol of toxin per mole of antibody was accomplished. The drug/Ab ratio was determined by peak area integration.

Zhang S., Zhou D., Zheng C., Xiong P., Zhu W, & Zheng D. (2021). Preclinical evaluation of a novel antibody-drug conjugate targeting DR5 for lymphoblastic leukemia therapy. Molecular Therapy Oncolytics, 21, 329-339.

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Quantifying Clindamycin in Whole Blood

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Whole blood was collected (200 μL) in an EDTA-K2 microtainer and was processed immediately prior to freezing at the study sites. PK samples were sent to the Pediatric Trials Network central laboratory (OpAns, LLC, Durham, NC) for storage and analysis. Clindamycin concentrations were quantified using a validated liquid chromatography-tandem spectrometry (LC-MS/MS) assay (assay did not quantify metabolite concentrations). The chromatography system and mass spectrometer used for sample analysis were the Agilent 1200 series high-performance liquid chromatography (HPLC) and an Agilent 6400 series triple quadrupole system, respectively. The Pursuit XRS Ultra C18 column (50 mm length x 2 mm internal diameter, 2.8 μm particle size, Agilent) and a gradient mobile phase (water containing 0.5% (v/v) formic acid; methanol containing 0.1% (v/v) formic acid) were used. The validation range for the assay was 50–50,000 ng/mL. The lower limit of quantification was 50 ng/mL. Accuracy and precision were within the FDA bioanalytical assay validation criteria (e.g., ±15%).

Gonzalez D., Melloni C., Yogev R., Poindexter B.B., Mendley S.R., Delmore P., Sullivan J.E., Autmizguine J., Lewandowski A., Harper B., Watt K.M., Lewis K.C., Capparelli E.V., Benjamin DK J.r, & Cohen-Wolkowiez M. (2014). Use of Opportunistic Clinical Data and a Population Pharmacokinetic Model to Support Dosing of Clindamycin for Premature Infants to Adolescents. Clinical pharmacology and therapeutics, 96(4), 429-437.

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Comprehensive Characterization of Molecular Structure

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Specific rotation was measured by a JASCO P-2000 polarimeter with a 10 mm cell at 20 °C. Ultraviolet (UV) data were acquired by an Applied Photophysics Chirascan-Plus circular dichroism spectrometer using a 1 mm UV cell. Infrared (IR) spectra were recorded by a JASCO Fourier transform/infrared spectrometer (FT/IR-4200). One-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectra were obtained by a Bruker Avance III HD 800 MHz NMR spectrometer located at the College of Pharmacy, Seoul National University, Republic of Korea. Chemical shifts of all NMR spectra were referenced to the residual protonated solvent peaks of dimethyl sulfoxide (DMSO)-d₆ (δ_H 2.50/δ_C 39.5). Liquid chromatography/mass spectrometry (LS/MS) data and low-resolution electrospray ionization mass spectrometry (LRESIMS) data were acquired using an Agilent Technologies 1200 series high-performance liquid chromatography (HPLC) coupled with an Agilent Technologies 6130 series single quadrupole ESIMS instrument. High-resolution ESIMS (HRESIMS) experiments were carried out on an AB Sciex 5600 quadrupole time-of-flight (QTOF) HRMS instrument at the National Instrumentation Center for Environmental Management (NICEM), Seoul National University, Republic of Korea.

Kang S., Han J., Jang S.C., An J.S., Kang I., Kwon Y., Nam S.J., Shim S.H., Cho J.C., Lee S.K, & Oh D.C. (2022). Epoxinnamide: An Epoxy Cinnamoyl-Containing Nonribosomal Peptide from an Intertidal Mudflat-Derived Streptomyces sp.. Marine Drugs, 20(7), 455.

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